Liliumpolysaccharide Extract Antioxidant Activity And Its Effect On Tumor Cell

Lily,as a edible and medicainal Chinese herbal medicine is treasure traditional Chinese medicine.polysaccharides as one of its active ingredient,has anti-oxidation,anti-tumor,anti-virus,anti-depressantimmunomodulatory,and othermultiple roles.Lily has many varieties,each of them all have different components and functions and focus on different side.Many of them have been proved to have ability of anti-oxidation and anti-tumor.As a competitive industries and High-quality type from Hunan province ShaoYang Longhui,Longya lilium has become a pillar industry in longhui county,also the national geographical indication products.But Longya lilium is few reported on the research,it have considerable developing potential.Based on the optimization of polysaccharide extraction process,explored it’s ability of anti-oxidation and anti-tumor to HepG2 cells,SKOV3 cells and HeLa cells.Choose two components toexplored by which the application of polysaccharide from Longya lilium will been broaden.1.Through to the extraction temperature,extraction solid-liquid ratio and extraction times four factors in the orthogonal experiment,the optimal process of lily’s polysaccharide extraction is:extraction temperature as 50℃,extraction time as 40min,solid-liquid as 1:7and extraction times quintic.lily’s polysaccharide extraction ratio can reach 8%2.By reviewing Sevag method,enzyme method,method of Enzyme—Severing,Eluting with organic agents and EAE-52 cellulose five different methods of deproteinization.Results showed that DEAE-52 cellulose can depigmentation and completely remove protein,make coarse polysaccharide purified about 4 times.Method of Enzyme—Severing removes protein effectively.Raise the rate of removal to 84.83%,control lossing rate of polysaccharid within 20%.3.To explore crude polysaccharide extract（LP₁）,Enzyme—Severing extract（LP₂）,organic agents extract（LP₃）and DEAE extract（LP₄）four Longya lilium extract hydroxyl free radical ability,remove superoxide free radical ability,antioxidant activities by DPPH assay.Results showed the antioxidant effect of four extracts all have certain antioxidant capacity.4.MTT assay was used to evaluate anti-tumor activities of LP₁、LP₂、LP₄.against HepG2cells、SKOV3 cells、HeLa cells.Results showed HepG2 cells were processced by LP₄solution effectively with concentration of 1mg/mL,inhibition rate were 46.36%,LP₂inhibition rate were 24.35%,LP₂ inhibition rate were 24.35%.LP₁ has best inhibition activity on SKOV3 cells,with concentration of 2mg/mL inhibition rate were 35.58%,LP₂ inhibition rate were17.72%、LP₄inhibition rate were 12.5%;three extract were have not effectively inhibition result on Hela cells.5.Annexin-V/PI double staining method and western blot for mechanism of apoptosis LP₁andLP₄on HepG2 cells and SKOV3 cells.Results showed both are late apoptotic rate higher than early apoptosis rate.Longya lilium extract can raised Bax protein expression,and cut Bcl-2 protein expression,activated Caspaes-3 of expression.The LP₁and LP₄ induce cell to apoptosisthough mitochondrial pathway and death receptor pathway.